Simultaneous detection of enteric bacteria from surface waters by QPCR in comparison with conventional bacterial indicators.
A rapid quantitative polymerase chain reaction (QPCR) method was developed for simultaneous detection of enteric bacteria from surface waters by utilizing a pair of universal primers which targeted four bacteria strains, namely Shigella dysenteriae, Vibrio cholerae, Salmonella typhimurium, and Escherichia coli. It was estimated that the QPCR method had a 94% confidence, and a detection limit as 2.7 E. coli cells per sample in undiluted DNA extracts. The QPCR method was applied for the bacteriological examination of several surface waters in the urban area of Xi'an, China and comparison was made with the conventional bacteria indicators determined by conventional membrane filter (MF) method. As a result, the calibrator cell equivalents (CCE) determined by QPCR was 2.2 to five times of the total coliform CFU, and the characteristics of the bacterial quality of different waters could be well presented by the QPCR results with a higher sensitivity. The coefficient of variation (CV) of data obtained by QPCR was smaller than that by traditional MF method, indicating a more stable analysis result. The QPCR method could thus be used as a supplement of the conventional culture method for more sensitive detection of pathogenic enteric bacteria from water.
Liu
Y J
YJ
Key Laboratory of Northwest Water Resource, Ecology and Environment, Ministry of Education, Xi'an University of Architecture and Technology, Xi'an, 710055, China. liuyongjun@xauat.edu.cn
Zhang
C M
CM
Wang
X C
XC
D001431:Bacteriological Techniques / Q000592:standards*; D002681:China; D004755:Enterobacteriaceae / Q000235:genetics* / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D005374:Filtration; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012486:Salmonella typhimurium / Q000235:genetics / Q000302:isolation & purification; D012762:Shigella dysenteriae / Q000235:genetics / Q000302:isolation & purification; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification; D014871:Water Microbiology
D012336:RNA, Ribosomal, 16S